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两种新晶体形式的嗜热栖热放线菌Cel7A的结晶及初步X射线衍射分析。

Crystallization and preliminary X-ray diffraction analysis of Hypocrea jecorina Cel7A in two new crystal forms.

作者信息

Bodenheimer Annette M, Cuneo Matthew J, Swartz Paul D, He Junhong, O'Neill Hugh M, Myles Dean A A, Evans Barbara R, Meilleur Flora

机构信息

Molecular and Structural Biochemistry Department, North Carolina State University, Raleigh, North Carolina, USA.

Biology and Soft Matter Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA.

出版信息

Acta Crystallogr F Struct Biol Commun. 2014 Jun;70(Pt 6):773-6. doi: 10.1107/S2053230X14008851. Epub 2014 May 10.

Abstract

Cel7A (previously known as cellobiohydrolase I) from Hypocrea jecorina was crystallized in two crystalline forms, neither of which have been previously reported. Both forms co-crystallize under the same crystallization conditions. The first crystal form belonged to space group C2, with unit-cell parameters a=152.5, b=44.9, c=57.6 Å, β=101.2°, and diffracted X-rays to 1.5 Å resolution. The second crystal form belonged to space group P6₃22, with unit-cell parameters a=b≃155, c≃138 Å, and diffracted X-rays to 2.5 Å resolution. The crystals were obtained using full-length Cel7A, which consists of a large 434-residue N-terminal catalytic domain capable of cleaving cellulose, a 27-residue flexible linker and a small 36-residue C-terminal carbohydrate-binding module (CBM). However, a preliminary analysis of the electron-density maps suggests that the linker and CBM are disordered in both crystal forms. Complete refinement and structure analysis are currently in progress.

摘要

来自嗜热栖热放线菌的Cel7A(以前称为纤维二糖水解酶I)以两种晶体形式结晶,这两种形式以前均未报道过。两种形式在相同的结晶条件下共结晶。第一种晶体形式属于空间群C2,晶胞参数为a = 152.5、b = 44.9、c = 57.6 Å,β = 101.2°,X射线衍射分辨率为1.5 Å。第二种晶体形式属于空间群P6₃22,晶胞参数为a = b ≃ 155,c ≃ 138 Å,X射线衍射分辨率为2.5 Å。这些晶体是使用全长Cel7A获得的,它由一个能够切割纤维素的434个残基的大N端催化结构域、一个27个残基的柔性连接子和一个36个残基的小C端碳水化合物结合模块(CBM)组成。然而,对电子密度图的初步分析表明,连接子和CBM在两种晶体形式中都是无序的。目前正在进行完整的精修和结构分析。

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